Positively regulated glycerol/G3P-dependent Bacillus subtilis gene expression system based on anti-termination.
Plasmid pLALA was constructed for glycerol or glycerol-3-phosphate inducible plasmid-borne gene expression in Bacillus subtilis and closely related Gram-positive bacteria. Gene expression using pLALA is based on anti-termination of transcription and involves the B. subtilis GlpP protein that in the presence of glycerol-3-phosphate acts as an anti-terminator protein by binding to the 5'-untranslated region of glpD mRNA. Properties and the usefulness of the system, denoted LALA, were validated by inducible production in B. subtilis strains of two water-soluble proteins (beta-galactosidase and a protein phospho-tyrosine phosphatase), and one integral membrane protein (heme A synthase). Advantages with LALA is that it is based on positive control, does not involve a DNA-binding protein, and that glycerol, a cheap and stable compound, can be used as inducer of gene expression.
Lewin
Anna
A
Department of Cell and Organism Biology, Lund University, Lund, Sweden.
Su
Xiao-Dong
XD
Hederstedt
Lars
L
D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D005990:Glycerol / Q000378:metabolism*; D005994:Glycerophosphates / Q000378:metabolism*; D010957:Plasmids; D017027:Protein Tyrosine Phosphatases / Q000096:biosynthesis / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D001426:Bacterial Proteins; D005994:Glycerophosphates; D016601:RNA-Binding Proteins; C064872:antiterminator proteins, Bacteria; C029620:alpha-glycerophosphoric acid; D017027:Protein Tyrosine Phosphatases; D001616:beta-Galactosidase; D005990:Glycerol